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Transcription factor for improving plant seed fatty acid content, coding gene and application thereof

A fatty acid and encoding technology, applied in the direction of plant genetic improvement, application, plant peptides, etc.

Active Publication Date: 2014-04-23
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, no specific technical publications have been published on the regulation of fatty acid content or composition by cotton transcription factors

Method used

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  • Transcription factor for improving plant seed fatty acid content, coding gene and application thereof
  • Transcription factor for improving plant seed fatty acid content, coding gene and application thereof
  • Transcription factor for improving plant seed fatty acid content, coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment 1, the acquisition of the coding gene GhDof1 of cotton transcription factor GhDof1

[0074] 1. Total RNA extraction: Upland cotton (Gossypium hirsutum L.) variety Coker201 (Wu Xiuming, Liu Chuanliang, Zhang Chaojun, Li Fuguang. Research progress in cotton somatic embryogenesis. Bulletin of Botany, 2008,25(4):469-475 ) as the experimental material, the immature embryo material 20 days after flowering was taken, quick-frozen in liquid nitrogen, and CTAB method was applied (references: Liu Yang, He Xinyao, Ma Hongbo, Wu Yongli, Yang Youming. The total RNA of each tissue of cotton was extracted by CTAB-PVP method Research. Journal of China Agricultural University, 2006, 11(1):53-56) to extract RNA.

[0075] 2. Use Promaga's RT-PCR reaction system to reverse transcribe the RNA extracted in step 1 to synthesize cDNA.

[0076] 3. RT-PCR amplification:

[0077] According to the EST information of cotton in the NCBI database, spliced ​​into contig, the following pri...

Embodiment 2

[0085] Example 2, Functional Analysis of Transcriptional Activation of GhDof1 Transcription Factor

[0086] In this example, pGBT9 (Meng Hongyan, Du Xiongming, Zhang Chunyi, etc. Cloning of cotton transcription factor gene GhMS3 and identification of its promoter function. Chinese Agricultural Sciences, 2010, 43 (17)) was used as the original vector to construct the following three expression vectors ( figure 2 Middle A): (1) the recombinant expression vector pGBT-GhDof1 containing the full-length CDS of the GDof1 gene (sequence 1, referred to as GDof1-CDS); (2) containing the N-terminal of the GDof1 gene (the 1-501st nuclear (GDof1-N for short) recombinant expression vector pGBT-GhDof1-N; (3) recombinant expression vector pGBT containing the C-terminal of the GhDof1 gene (nucleotides 481-948 of sequence 1, GhDof1-C for short) -GhDof1-C. In order to analyze the transcription activation function of GhDof1 transcription factor.

[0087] 1. Construction of recombinant express...

Embodiment 3

[0102] Embodiment 3, expression analysis of GhDof1 gene

[0103] RNA was extracted from the ovary, anther, calyx, petal, ovule, root, stem and leaf tissue of upland cotton (Gossypium hirsutum L.) variety Coker201, and cDNA was synthesized by reverse transcription with Promaga's RT-PCR reaction system, and designed for the GhDof1 gene Primers 5'-TGATGTGAGTGAACCGAAGT-3' (positions 507-526 of Sequence 1) and 5'-TTTACGGGAGATAGAGGC-3' (positions 2-18 are the reverse complement of positions 932-948 of Sequence 1) to Ghactin As an internal reference gene, primers 5'-ATCCTCCGTCTTGACCTTG-3' and 5'-TGTCCGTCAGGCAACTCAT-3' were designed, and the expression of GhDof1 gene in different tissue parts was analyzed by semi-quantitative RT-PCR.

[0104] The result is as Figure 4 As shown, the GhDof1 gene exhibits certain tissue specificity, which is specifically manifested in: the gene is expressed in ovary, anther, calyx and other reproductive organ tissues, while the expression level in peta...

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Abstract

The invention discloses a transcription factor associated for improving plant seed fatty acid content, a coding gene and application thereof. The transcription factor provided by the invention is (a) or (b): (a) any protein segment with length of 155th to 315th amino acids which is obtained by extending the amino acid sequences of 161th to 315th sites with carboxyl terminals containing at least sequence 2 in a sequence table from 161th site in the sequence 2 towards an amino acid terminal of the sequence 2 according to the direction of the amino acid sequence of the sequence 2, wherein the protein segment has transcription activity; and (b) protein which is obtained by substituting, deleting and / or adding one or more amino acid residues for the amino acid sequence defined by (a) and has transcription activity or is associated for improving the plant seed fatty acid content and is derived from the amino acid sequence defined by (a). According to the transcription factor GhDof1 provided by the invention, the fatty acid content of plant seeds can be obviously increased; and the transcription factor is good in application prospect.

Description

technical field [0001] The invention relates to a transcription factor related to increasing the fatty acid content of plant seeds, its encoding gene and application, in particular to a transcription factor derived from cotton and related to increasing the fatty acid content of plant seeds, its encoding gene and application. Background technique [0002] Cotton is an important economic crop and oil crop in my country. my country has 6 million hectares of cotton planted all the year round, with an output of about 7.6 million tons of lint and 13 million tons of cottonseed. As the main by-product of cotton production, cottonseed is rich in oil and protein (Xu Hongxia, Yang Weihua, Wang Yanqin, Zhou Dayun, Kuang Meng, Feng Xinai. Analysis of the quality of cottonseed for oil in my country. China Cotton, 2009,36(7):2-3) , the fat content of cottonseed after shelling is 30% to 40%, which can produce oil. In recent years, with the increasing shortage of oil resources and energy re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N5/10C12N1/21C12N1/19C12N15/82C12N15/84A01H5/00
Inventor 华金平刘正杰张园赵鹏赵清翠李玉华王玉美
Owner CHINA AGRI UNIV